本文選題：線粒體 + 頭發(fā)��； 參考：《重慶醫(yī)科大學(xué)》2006年碩士論文

【摘要】： 目的:研究頭發(fā)mtDNA4977bp片段缺失與人群年齡的關(guān)系,建立相關(guān)方程,以期在微量物證中尋找更多的信息,為分子水平構(gòu)建犯罪分子特征提供更多的可能。 方法:按中國(guó)人口自然分布規(guī)律,隨機(jī)剪取54名漢族健康志愿者的頭發(fā)若干,各取4根用無(wú)水乙醇清洗、蒸餾水漂洗后,自然風(fēng)干,從近毛囊部位截取1.5cm,剪成0.3cm左右的片段,放入0.5ml EP管中,加chelex-100和蛋白酶K消化抽提mtDNA,用所設(shè)計(jì)的引物分別對(duì)mtDNA保守區(qū)和4977bp缺失突變區(qū)進(jìn)行聚合酶鏈反應(yīng)(PCR),若4977bp處不發(fā)生缺失,常規(guī)PCR不能擴(kuò)增正常約5000bp的堿基,所以沒(méi)有產(chǎn)物,反之,可以擴(kuò)增出線粒體片段缺失后相應(yīng)遺留序列的PCR產(chǎn)物。將擴(kuò)增產(chǎn)物經(jīng)瓊脂糖凝膠電泳分離,EB染色后,在Bio-Rad紫外凝膠成像儀掃描成像,確定其激光密度。以保守內(nèi)參照區(qū)擴(kuò)增產(chǎn)物激光密度作為總量,缺失區(qū)擴(kuò)增產(chǎn)物激光密度值作為缺失量,用缺失量/總量計(jì)算缺失率。 結(jié)果:25歲以下幾乎無(wú)缺失,缺失區(qū)很少擴(kuò)增出產(chǎn)物,大于25歲者缺失區(qū)逐漸可擴(kuò)增出產(chǎn)物,且隨年齡增長(zhǎng),相同條件下擴(kuò)增產(chǎn)物增多,4977bp缺失率增大,各年齡段組間比較均P0.05。根據(jù)統(tǒng)計(jì)分析,缺失率與年齡有直線相關(guān)趨勢(shì),相關(guān)系數(shù)r=0.98,可以建立相關(guān)回歸方程Age=0.74(±0.02)×dmtDNA+21.03(±0.80)(dmtDNA為缺失
[Abstract]:Objective: To study the relationship between the loss of mtDNA4977bp fragments of hair and the age of the population and establish the correlation equation, in order to find more information in the trace evidence, and to provide more possibilities for the construction of criminal characteristics at the molecular level.
Methods: according to the natural distribution law of Chinese population, the hair of 54 Han healthy volunteers was cut randomly, each 4 was washed with anhydrous ethanol. After the distilled water was rinsed, the natural wind was dried, the 1.5cm was cut from the near hair follicle, and the fragments were cut into about 0.3cm. In the 0.5ml EP tube, the mtDNA was digested with Chelex-100 and protease K, and the designed citation was used. Polymerase chain reaction (PCR) was performed on the mtDNA conservative region and the 4977bp deletion mutation region respectively. If no deletion occurred at 4977bp, the conventional PCR could not amplify the normal base of approximately 5000bp, so there was no product. On the contrary, the PCR products of the corresponding remnants of the mitochondrial fragment could be amplified. The amplified products were separated by agarose gel electrophoresis and EB. After staining, the laser density was determined by scanning the Bio-Rad ultraviolet gel imaging instrument. The laser density of the amplified product in the internal reference area was taken as the total amount, the laser density value of the amplified product in the missing region was used as the missing amount, and the loss rate was calculated by the amount of loss / total.
Results: there was almost no deletion under 25 years of age, and there was little amplification of the product in the missing area, and the product was gradually amplified in the missing area of the people over 25 years old, and with the increase of age, the amplification products increased and the loss rate of 4977bp increased. The correlation coefficient of the loss rate was linearly correlated with age, and the correlation coefficient was r=0.98, as compared with the statistical analysis of P0.05.. Correlation regression equation Age=0.74 (+ 0.02) x dmtDNA+21.03 (+ 0.80) can be established (dmtDNA is missing).
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：D919

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本文編號(hào)：1995019